O-acyloxime derivatives, preparation method thereof, and pharmaceutical composition containing the same for prevention and treatment of cardiovascular disease

ABSTRACT

The present invention relates to novel O-acyloxime derivatives, a preparation method thereof and a pharmaceutical composition comprising the same for prevention and treatment of cardiovascular disease. 
     The O-acyloxime derivatives according to the present invention may valuably be used for prevention and treatment of cardiovascular diseases such as hyperlipidemia, coronary arterial heart disease, atherosclerosis, and myocardial infarction caused by Lp-PLA 2 , because they have excellent inhibitory effect of Lp-PLA 2 .

CROSS-REFERENCE TO RELATED APPLICATIONS

This patent application is a Divisional of U.S. patent application Ser.No. 11/163,383, filed on Oct. 17, 2005, which in turn claims the benefitof priority from Korean Patent Application No. 10-2004-0092263, filed onNov. 12, 2004, the contents of each of which are incorporated herein byreference.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates to novel O-acyloxime derivatives, apreparation method thereof and a pharmaceutical composition comprisingthe same for prevention and treatment of cardiovascular disease.

2. Description of the Prior Art

Recently, mortality from coronary heart disease (CHD) is significantlyincreased, and atherosclerosis is one of the major causes of death.Atherosclerosis is an inflammatory disease caused by accumulation oflipid and fibrin on the arterial wall, and the major causes of thedisease are hypertension, smoking, obesity, increase of low-densitylipoprotein (LDL) in plasma. However, another cause of theatherosclerosis may be inferred from the fact that more than 50% ofpatients have contracted atherosclerosis regardless of the above causes.According to the investigation result of 580 patients having coronaryheart disease and 1,160 normal men, which is reported by the West ofScotland Coronary Prevention Study (WOSCOPS), lipoprotein-associatedPhospholipase A₂(Lp-PLA₂) levels of the patients were significantly highcompared to that of the normal men (N. Engl. J. Med., 2000, 343,1148-1155), and it has been identified that Lp-PLA₂ is an independentrisk factor of coronary heart disease (Expert Rev. Mol. Diagn., 2002, 2,17-22).

The molecular weight of Lp-PLA₂ is 45 kDa. Lp-PLA₂ is a secretedcalcium-independent member type VII of phospholiphase A₂ superfamilymainly formed by monocyte, macrophage, T-lymphocytes, and mast cells,and is known as an enzyme of platelet-activating factor acetylhydrolase(PAF-AH, EC 3.1.1.47) (Arterioscler. Thromb. Vasc. Biol., 1996, 16,591-595). Additionally, 80% of Lp-PLA₂ is bound to LDL, and theremainder is bound to high-density lipoprotein (HDL) and verylow-density lipoprotein (VLDL) (Arterioscler. Thromb. Vasc. Biol., 1995,15, 1764-1773).

Accumulation of LDL, particularly oxidized LDL, on the arterial wall isknown as the most important initial step of atherosclerosis. Lp-PLA₂ isbound to LDL in a latent state until LDL is oxidized or modified. As LDLis oxidized, Lp-PLA₂ is activated and forms a large amount oflysophosphatidylcholine (lyso-PC) and free oxidized fatty acids byrapidly hydrolyzing the sn-2 fatty acid of oxidized phospholipid(Biochem. J., 1999, 338, 479-487). LDL is oxidized in intima and servesas a substrate of Lp-PLA₂. Hydrolyzed products further acceleratechronic inflammation related to accumulation of macrophage, and apositive feedback mechanism of macrophage forming a large amount ofLp-PLA₂ further accelerates progress of vascular disorder. Biologicalactivity has not been defined completely yet, because structures of freeoxidized fatty acids formed by Lp-PLA₂ are not clearly identified. Fattyacids of micromolar concentration formed from ox-LDL are biologicallyinactive. However, in 1990s, reports on pro-inflammatory andpro-atherogenic role of lyso-PC, which is another decomposed product,were rapidly increased. For example, the reported roles are impairmentof endothelium-dependent relaxation, inducement of vascular cell andintracellular adhesion molecules, activity as chemoattractant ofmonocyte and T-lymphocytes, suppressed production and release ofendothelium-derived nitric oxide, inhibition of macrophage migration,toxicity at the concentration higher than 30-50 micromole, and releasestimulation of arachidonic acid from endothelial cells (Curr. Opin.Pharmacol., 2001, 1, 121-125).

Lp-PLA₂ is an independent risk factor of coronary artery disease in thecase of hypercholestrolemia patients, and is suggested as apro-inflammatory agent, which is detected in macrophage ofatherosclerotic lesions (Arterioscler. Thromb. Vasc. Biol., 1999, 19,2909-2971). According to recent researches, the formation of fattystreak in Watanabe heritable hyperlipidemic rabbits, which is a modelanimal of atherosclerosis, is significantly decreased by dosing Lp-PLA₂inhibitor (Atherosclerosis, 2000, 151, 166). Therefore, inhibition ofLp-PLA₂ activity is highlighted as a target for prevention and treatmentof atherosclerosis (N. Engl. J. Med., 2000, 343, 1148-55). Accordingly,research and development of Lp-PLA₂ inhibitor will be very important forprevention and treatment of atherosclerosis.

GlaxoSmithKline isolated a series of novel inhibitors of Lp-PLA₂ fromthe culture broths of Pseudomonas fluorescens (J. Antibiotics, 2000, 53,664-669). A potent, orally active Lp-PLA₂ inhibitor, SB-480848, has beendeveloped from their synthetic derivatives, and SB-480848 is currentlyin a phase II clinical study.

Through researches on novel drugs for treatment of hyperlipidemia andatherosclerosis, the inventors have synthesized O-acyloxime derivatives,and completed the present invention by identifying that these compoundshave inhibitory effects on Lp-PLA₂.

SUMMARY OF THE INVENTION

The present invention provides novel O-acyloxime derivatives.

Additionally, the present invention provides a preparation method of theO-acyloxime derivatives.

Additionally, the present invention provides a pharmaceuticalcomposition for prevention and treatment of cardiovascular disease,comprising the O-acyloxime derivatives as active ingredients.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides novel O-acyloxime derivatives representedby the following Formula 1:

Wherein:

R¹ is aryl; heteroaryl such as furan, thiophene, imidazole, pyrrole,pyridine, pyrimidine and so on; aryl or heteroaryl substituted by one ormore group selected from halogen, C₁˜C₆ alkyl, C₁˜C₆ alkoxy, C₁˜C₆alkylthio, cyano, amino, mono- or di-C₁˜C₆ alkylamino, C₂˜C₆alkylcarbonyl, C₁˜C₆ alkyl substituted by one or more halogen, aryl,C₆˜C₁₀ arylalkyl, aryl or C₆˜C₁₀ arylalkyl substituted by one or morehalogen, and C₆˜C₁₀ arylcarbonyl; C₂˜C₆ heterocyclic ring comprising oneor more N, O or S such as tetrahydrofuran, tetrahydropyran, piperidine,morpholine, thiomorpholine and so on; arylalkenyl;

R² is hydrogen; halogen; C₁˜C₆ alkyl; hydroxy; amino; cyano; C₁˜C₆alkylamino; di(C₁˜C₆ alkyl)amino; (C₁˜C₃)alkylcarbonylamino; di(C₁˜C₃alkylcarbonyl)amino; C₁˜C₆ alkoxy; aryl(C₁˜C₃)alkoxy; heteroaryl such aspyridine and pyrimidine; C₂˜C₆ heterocyclic ring comprising one or moreN, O or S such as tetrahydrofuran, tetrahydropyran, piperidine,morpholine, thiomorpholine and so on; and

R³ is aryl; heteroaryl such as furan, thiophene, imidazole, pyrrole,pyridine, pyrimidine and so on; aryl or heteroaryl substituted by one ormore halogen, nitro, amino, C₁˜C₆ alkyl, and C₁˜C₆ alkyl substituted byone or more halogen; C₂˜C₆ heterocyclic ring comprising one or more N, Oor S such as tetrahydrofuran, tetrahydropyran, piperidine, morpholine,thiomorpholine and so on; C₁˜C₁, alkyl; C₁˜C₁, alkenyl; C₃˜C₁₀cycloalkyl; mono- or di-C₁˜C₆ alkylamino; C₁˜C₆ alkylcarbonylamino;di(C₁˜C₆ alkylcarbonyl)amino; arylcarbonylamino; di(arylcarbonyl)amino.

Preferably,

R¹ is phenyl; heteroaryl such as furan, thiophene, pyrrole; phenylsubstituted by one or more group selected from halogen, C₁˜C₆ alkyl,C₁˜C₆ alkoxy; styrene;

R² is hydrogen; halogen; C₁˜C₆ alkyl; amino; cyano; di(C₁˜C₃alkylcarbonyl)amino; benzyloxy; and

R³ is phenyl; heteroaryl such as furan and thiophene; phenyl substitutedby one or more halogen, nitro; morpholine; C₁˜C₁₈, alkyl; C₁˜C₁₈alkenyl; cyclohexane; benzoylamino.

Further preferred compounds among the O-acyloxime derivatives accordingto the present invention are listed as follows, and their structures areshown in Table 1.

-   1) (E)-benzaldehyde-O-benzoyloxime,-   2) (E)-benzaldehyde-O-morpholine-4-carbonyloxime,-   3) (E)-benzaldehyde-O-cyclohexanecarbonyloxime,-   4) (E)-benzaldehyde-O-benzoylthiocarbamic acid oxime-   5) (E)-benzaldehyde-O-thiophene-2-carbonyloxime,-   6) (E)-benzaldehyde-O-furan-2-carbonyloxime,-   7) (E)-4-fluorobenzaldehyde O-benzoyloxime,-   8) (E)-4-fluorobenzaldehyde O-4-fluorobenzoyloxime,-   9) (E)-4-fluorobenzaldehyde O-morpholine-4-carbonyloxime,-   10) (E)-(3,5-di-t-butyl)-4-methoxybenzaldehyde O-benzoyloxime,-   11) (E)-3-phenylpropenaldehyde O-benzoyloxime,-   12) (E)-1-amino-benzaldehyde O-benzoyloxime,-   13) (E)-1-N,N-diacetylamino-benzaldehyde O-benzoyloxime,-   14) (E)-1-cyano-benzaldehyde O-benzoyloxime,-   15) (E)-1-chloro-benzaldehyde O-benzoyloxime,-   16) (E)-1-benzoic acid-benzaldehyde O-benzoyloxime,-   17) (E)-2-furan-2-carboaldehyde O-benzoyloxime,-   18) (E)-2-thiophene-2-carboaldehyde O-benzoyloxime,-   19) (E)-1-phenylethanonaldehyde O-benzoyloxime,-   20) (E)-N-4-fluorobenzylpyrrole-2-carboaldehyde O-benzoyloxime,-   21) (E)-3,4-di-fluorobenzaldehyde O-benzoyloxime,-   22) (E)-3,4-di-fluorobenzaldehyde O-morpholine-4-carbonyloxime,-   23) (E)-3,4-di-fluorobenzaldehyde O-4-fluorobenzoyloxime,-   24) (E)-3,4-di-fluorobenzaldehyde O-oleyloxime,-   25) (E)-3,4-di-fluorobenzaldehyde O-linoleyloxime,-   26) (E)-3,4-di-fluorobenzaldehyde O-decanoyloxime,-   27) (E)-3,4-di-fluorobenzaldehyde O-4-nitro)benzoyloxime,-   28) (E)-3,4-di-fluorobenzaldehyde O-3,4-difluorobenzoyloxime,-   29) (E)-2,3-di-fluorobenzaldehyde O-benzoyloxime,-   30) (E)-2,3-di-fluorobenzaldehyde O-morpholine-4-carbonyloxime,-   31) (E)-2,4-di-fluorobenzaldehyde O-benzoyloxime,-   32) (E)-2,4-di-fluorobenzaldehyde O-morpholine-4-carbonyloxime,-   33) (E)-2,4-di-fluorobenzaldehyde O-4-fluorobenzoyloxime,-   34) (E)-2,6-di-fluorobenzaldehyde O-benzoyloxime,-   35) (E)-2,6-di-fluorobenzaldehyde O-morpholine-4-carbonyloxime,-   36) (E)-3,5-di-fluorobenzaldehyde O-benzoyloxime,-   37) (E)-3,5-di-fluorobenzaldehyde O-morpholine-4-carbonyloxime, and-   38) (E)-2,5-di-fluorobenzaldehyde O-morpholine-4-carbonyloxime.

TABLE 1 Compound Structure 1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

27

28

29

30

31

32

33

34

35

36

37

38

The O-acyloxime derivatives represented by Formula 1 may be used asforms of pharmaceutically acceptable salts thereof, and pharmaceuticallyacceptable free acids salts thereof are preparable. Both organic andinorganic acids may be used as the free acids. For example, theinorganic acids include hydrochloric acid, hydrobromic acid, sulfuricacid, and phosphoric acid, and the organic acids include citric acid,acetic acid, lactic acid, maleic acid, umaric acid, gluconic acid,methanesulfonic acid, glyconic acid, succinic acid, 4-toluenesulfonicacid, trifluoroacetic acid, galutronic acid, embonic acid, glutamicacid, and aspartic acid.

Additionally, the present invention provides a preparation method ofO-acyloxime derivatives represented by the following Chemical Reactionscheme 1.

wherein, R¹, R², and R³ are same as defined in Formula 1.

A preparation method for O-acyloxime derivatives according to thepresent invention contains the steps of:

1) obtaining a compound 3 by reacting a compound 2 with hydroxyamineunder the presence of base, and

2) obtaining a compound 1 by reacting the compound 3 prepared in thestep 1) with acylchloride.

The preparation method of O-acyloxime derivatives according to thepresent invention will be described in more detail as follows.

In the step 1, the base includes metal hydroxides such as sodiumhydroxide, potassium hydroxide, and lithium hydroxide; basic salts suchas sodium carbonate, potassium carbonate, cesium carbonate, sodiumbicarbonate, and sodium acetate; aromatic amines such as pyridine andlutidine; and tertiary amines such as triethylamine, tripropylamine,tributylamine, cyclohexyldimethylamine, 4-dimethylaminopyridine,N,N-dimethylaniline, N-methylpiperidine, N-methylpyrrolidine, andN-methylmorpholine. Triethylamine is preferable.

Water, C1˜C3 low alcohol, or a mixture thereof is preferably used as areaction solvent, and ethanol is most preferably used.

Reaction temperature is preferably room temperature. Reaction time ispreferably 30 min˜24 hrs, and is most preferably 1˜20 hrs.

Additionally, the compound 3 may be formed as a mixture of isomers, andthe isomers may be separated by conventional methods such as columnchromatography.

In the step 2, the reaction may be performed either under the presenceor absence of a base. The base includes metal hydroxides such as sodiumhydroxide, potassium hydroxide, lithium hydroxide; basic salts such assodium carbonate, potassium carbonate, cesium carbonate, sodiumbicarbonate, and sodium acetate; aromatic amines such as pyridine andlutidine; and tertiary amines such as triethylamine, tripropylamine,tributylamine, cyclohexyldimethylamine, 4-dimethylaminopyridine,N,N-dimethylaniline, N-methylpiperidine, N-methylpyrrolidine, andN-methylmorpholine. Triethylamine or sodium hydride is preferable.

Additionally, sulfuric acid (H₂SO₄) may be used as a catalyst in thereaction, and dichloromethane, THF, or acetyl anhydride is preferablyused as a reaction solvent.

The reaction temperature is preferably about −5° C.˜+5° C., and is mostpreferably 0° C. The compound 3 may be prepared by a method disclosed inthe prior art or a similar method thereto, or is a commerciallyavailable compound may also be used.

Additionally, the O-acyloxime derivative of the compound 1 prepared inthe above may be formed as a mixture of isomers, and the isomers may beseparated by conventional methods such as column chromatography.

Additionally, the O-acyloxime derivative of the compound 1 may beprepared by reaction of the compound 2 of more than 2 equivalents withNH₂OH under the presence of a base, without passing through the step 2.

Additionally, the present invention provides a pharmaceuticalcomposition for prevention and treatment of cardiovascular disease,containing O-acyloxime derivatives of Formula 1 as active ingredients.

O-Acyloxime derivatives according to the present invention haveexcellent inhibitory effect on Lp-PLA₂, and thereby may effectively beused for prevention and treatment of cardiovascular disease caused byLp-PLA₂ such as hyperlipidemia, coronary arterial heart disease,atherosclerosis, and myocardial infarction.

The composition according to the present invention may further includeone or more active ingredients having the same or similar function inaddition to the O-acyloxime derivatives.

For the prevention and treatment of cardiovascular disease, thecomposition according to the present invention may be used independentlyor together with an operation, hormone treatment, pharmaceuticaltreatment and biological reaction controller.

The composition according to the present invention may be prepared byadding one or more pharmaceutically acceptable carriers in addition tothe above active ingredients. The pharmaceutically acceptable carriersinclude saline solution, sterile water, Ringer's solution, bufferedsaline solution, dextrose solution, maltodextrin solution, glycerol,ethanol, and a mixture thereof, and may further include otherconventional additives such as antioxidant, buffer solution, andbacteriostat, if necessary. Additionally, the composition may beformulated in various forms of injection such as solution, suspension,and emulsion; pill, capsule, granule, or tablet by further addingdiluent, dispersant, surfactant, bonding agent, and lubricant.Furthermore, the composition may be formulated with a proper methodknown in the art or a method disclosed by Remington's PharmaceuticalScience (the latest edition, Mack Publishing Company, Easton Pa.),according to the component or kinds of disease.

The composition according to the present invention may be prepared fororal administration, parenteral administration such as intravenous,subcutaneous, intraperitoneal administration, or local application. Thedosage varies according to the weight, age, sex, and health condition ofpatients, diet pattern, dosing intervals, dosing method, excretion rate,and state of disease. Daily dosage of O-acyloxime derivatives of Formula1 is about 0.1˜100 mg/kg, preferably 0.5˜10 mg/kg, and it is morepreferable to dose one to several times a day.

According to the result of toxicity test carried out by oraladministration of the O-acyloxime derivative according to the presentinvention, lethal dose 50 (LD₅₀) of the O-acyloxime derivative is morethan 1,000 mg/kg.

DETAILED DESCRIPTION OF EXEMPLARY EMBODIMENTS OF THE INVENTION

Hereinafter, preferred example embodiments of the present invention willbe described more fully for easier understanding. This invention may,however, be embodied in many different forms and should not be construedas limited to the example embodiments set forth herein.

Example 1 Preparation of (E)-Benzaldehyde O-Benzoyloxime

After dissolving benzaldehyde (5 ml, 49 mmol) in ethanol (100 ml),hydroxyamine (NH₂OH) (4.4 g, 64.0 mmol) and triethylamine (9 ml, 64.0mmol) were added. After stirring reaction solution for 1 hour at roomtemperature, the solvent was removed. After adding water (50 ml), thesolution was extracted with ethylacetate. The obtained residue was thenseparated into (E)-oxime compound (3 g, 50%) and (Z)-oxime compound(0.32 g, 5.4%) by column chromatography.

After dissolving the obtained (E)-oxime compound (0.22 g, 1.8 mmol) indichloromethane, benzoylchloride (0.3 ml, 2.3 mmol) was slowly added at0° C. under the presence of triethylamine. After stirring the reactionsolution for 1 hour, the reaction was quenched by 1N hydrochloric acid,and the reaction solution was extracted with dichloromethane. A purecompound of (E)-benzaldehyde O-benzoyloxime 1 (0.26 g, 65%) was obtainedby separating residue with silica gel column chromatography.

Additionally, after dissolving the obtained (Z)-oxime compound (0.1 g,0.83 mmol) in dichloromethane, reaction solution was cooled to −40° C.,and benzoylchloride (0.12 ml, 1.07 mmol) was added. After stirring for20 minutes, a compound of (E)-benzaldehyde O-benzoyloxime (1) (0.11 g,61%) was obtained by the same method as that of the (E)-oxime compoundpreparation.

¹H NMR (300 MHz, CDCl₃) 7.48 (m, 5H), 7.62 (m, 1H), 7.82 (dd, J=1.8, 7.8Hz, 2H), 8.14 (dd, J=2.1, 8.4 Hz, 2H), 8.57 (s, 1H).

Example 2 Preparation of (E)-benzaldehyde O-morpholine-4-carbonyloxime

After dissolving benzaldehyde (1.0 ml, 9.8 mmol) in ethanol (20 ml),hydroxyamine (0.88 g, 12.8 mmol) and triethylamine (1.77 ml, 12.8 mmol)were added. After stirring reaction solution for 1 hour at roomtemperature, the solvent was removed. After adding water (50 ml), thesolution was extracted with ethylacetate. (E)- and (Z)-oxime mixtures(1.00 g, 85%) were obtained by separating the residue with columnchromatography.

After dissolving the (E)- and (Z)-oxime mixtures (1.0 g, 8.26 mmol) inTHF (30 ml), morpholinecarbonylchloride (1.23 ml, 10.7 mmol) was slowlyadded at 0° C. under the presence of NaH (0.26 g, 10.7 mmol). Afterstirring the reaction solution for 1 hour, the reaction was quenched by1 N hydrochloric acid, and the reaction solution was extracted withdichloromethane. A pure compound of (E)-benzaldehydeO-morpholine-4-carbonyloxime (2) (1.8 g, 93%) was obtained by separatingthe residue with silica gel column chromatography.

¹H NMR (300 MHz, CDCl₃) 3.57 (t, J=4.9 Hz, 4H), 3.71 (t, J=4.6 Hz, 4H),7.45 (m, 3H), 7.73 (d, J=7.8 Hz, 2H), 8.31 (s, 1H)

Example 3 Preparation of (E)-Benzaldehyde-O-cyclohexanecarbonyloxime

The title compound was prepared in the same method as Example 1 exceptthat cyclohexanecarbonylchloride was used instead of benzoylchloride.

¹H NMR (300 MHz, CDCl₃) 2.0˜1.21 (m, 9H), 2.45 (m, 1H), 7.41 (m, 3H),7.72 (dd, J=1.8, 7.8 Hz, 2H), 8.34 (s, 1H).

Example 4 Preparation of (E)-Benzaldehyde-O-benzoylthiocarbamic acidoxime

The title compound was prepared in the same method as Example 1 exceptthat benzoylisocyanate was used instead of benzoylchloride.

¹H NMR (300 MHz, CDCl₃) 7.46 (m, 6H), 7.60 (m, 1H), 7.80 (m, 2H), 8.12(m, 2H), 8.55 (s, 1H).

Example 5 Preparation of (E)-benzaldehyde-O-thiophene-2-carbonyloxime

The title compound was prepared in the same method as Example 1 exceptthat thiophene-2-carbonylchloride was used instead of benzoylchloride.

¹H NMR (300 MHz, CDCl₃) 7.16 (dd, J=3.6, 4.8 Hz, 1H), 7.48 (m, 3H), 7.63(dd, J=1.2, 5.1 Hz, 1H), 7.81 (dd, J=1.8, 7.8 Hz, 2H), 7.94 (dd, J=1.2,4.2 Hz, 1H), 8.51 (s, 1H).

Example 6 Preparation of (E)-benzaldehyde-O-furan-2-carbonyloxime

The title compound was prepared in the same method as Example 1 exceptthat furan-2-carbonylchloride was used instead of benzoylchloride.

¹H NMR (300 MHz, CDCl₃) 6.54 (q, J=1.8 Hz, 1H), 7.30 (d, J=3.6 Hz, 1H),7.45 (m, 3H), 7.63 (s, 1H), 7.76 (t, J=4.1 Hz), 8.50 (s, 1H).

Example 7 Preparation of (E)-4-fluorobenzaldehyde O-benzoyloxime

The title compound was prepared in the same method as Example 1 exceptthat 4-fluorobenzaldehyde was used instead of benzaldehyde.

¹H NMR (300 MHz, CDCl₃) 7.14 (t like, J=8.7 Hz, 2H), 7.49 (t like, J=7.2Hz, 2H), 7.61 (t like, J=7.7 Hz, 1H), 7.82 (dd, J=5.4, 8.4 Hz, 2H), 8.12(d, J=7.2 Hz, 2H), 8.53 (s, 1H).

Example 8 Preparation of (E)-4-fluorobenzaldehyde O-4-fluorobenzoyloxime

The title compound was prepared in the same method as Example 1 exceptthat 4-fluorobenzaldehyde and 4-fluorobenzoylchloride were used insteadof benzaldehyde and benzoylchloride.

¹H NMR (300 MHz, CDCl₃) 7.15 (m, 4H), 7.80 (dd, J=5.3, 8.9 Hz, 2H), 8.12(dd, J=5.4, 8.4 Hz, 2H), 8.50 (s, 1H)

Example 9 Preparation of (E)-4-fluorobenzaldehydeO-morpholine-4-carbonyloxime

The title compound was prepared in the same method as Example 2 exceptthat 4-fluorobenzaldehyde was used instead of benzaldehyde.

¹H NMR (300 MHz, CDCl₃) 3.55 (t, J=4.9 Hz, 4H), 3.69 (t, J=4.6 Hz, 4H),7.08 (t, J=8.6 Hz, 2H), 7.71 (dd, J=5.5, 9.1 Hz, 2H), 8.27 (s, 1H).

Example 10 Preparation of (E)-(3,5-di-t-butyl)-4-methoxybenzaldehydeO-benzoyloxime

The title compound was prepared in the same method as Example 1 exceptthat 3,5-di-t-butylbenzoylaldehyde was used instead of benzoylchloride.

¹H NMR (300 MHz, CDCl₃) 1.44 (s, 18H), 3.72 (s, 3H), 7.44 (m, 5H, 7.66(s, 2H), 8.51 (s, 1H).

Example 11 Preparation of (E)-3-phenylpropenaldehyde O-benzoyloxime

The title compound was prepared in the same method as Example 1 exceptthat 3-phenylpropenaldehyde was used instead of benzaldehyde.

¹H NMR (300 MHz, CDCl₃) 7.11 (m, 2H), 7.40 (m, 3H), 7.50 (m, 3H), 7.60(m, 2H), 8.13 (m, 2H), 8.35 (dd, J=7.2, 7.8 Hz, 1H).

Example 12 Preparation of (E)-1-amino-benzaldehyde O-benzoyloxime

After dissolving benzonitrile (10.3 ml, 100 mmol) in ethanol (50 ml),hydroxyamine (9.02 g, 130 mmol) and sodium carbonate (13.76 g, 130 mmol)were added. After refluxing reaction solution for 6 hours, hydroxyamine(9.02 g, 130 mmol) and sodium carbonate (13.76 g, 130 mmol) were added,and the reaction solution was further refluxed for 14 hours. Afterremoving the solvent, water (50 ml) was added and the solution wasextracted with ethylacetate. (E)- and (Z)-oxime mixtures (12.6 g, 92.5%)were obtained by separating the residue with column chromatography.

After dissolving the (E)- and (Z)-oxime mixtures (0.73 g, 5.36 mmol) inCH₂Cl₂ (30 ml), benzoylchloride (0.81 ml, 6.79 mmol) was slowly added at0° C. under the presence of triethylamine (0.97 ml, 6.79 mmol). Afterstirring the reaction solution for 1 hour, there action was quenched by1 N hydrochloric acid, and the reaction solution was extracted withdichloromethane. A pure compound of (E)-1-amino-benzaldehydeO-benzoyloxime (12) (0.98 g, 75.9%) was obtained by separating theresidue with silica gel column chromatography.

¹H NMR (300 MHz, CDCl₃) 5.35 (s, 2H, —NH2), 7.47˜7.34 (m, 5H), 7.58 (m,1H), 7.72 (m, 2H), 8.05 (s, 2H).

Example 13 Preparation of (E)-1-N,N-diacetylamino-benzaldehydeO-benzoyloxime

After dissolving the (E)-1-amino-benzaldehyde O-benzoyloxime (0.2 g,0.83 mmol) obtained from Example 12 in acetic anhydride (10 ml), H₂SO₄(0.1 ml, catalystic amount) was added. After stirring reaction solutionfor 6 hours, water was added and the reaction solution was extractedwith dichloromethane. A pure compound of(E)-1-N,N-diacetylamino-benzaldehyde O-benzoyloxime (13) (0.23 g, 85%)was obtained by separating the residue with silica gel columnchromatography.

¹H NMR (300 MHz, CDCl₃) 2.44 (s, 6H), 7.50 (m, 5H), 7.63 (m, 1H), 7.88(m, 2H), 7.99 (m, 2H).

Example 14 Preparation of (E)-1-cyano-benzaldehyde O-benzoyloxime

After dissolving 2-hydroxyimino-2-phenylacetonitrile (1.0 g, 6.84 mmol)in CH₂Cl₂ (30 ml), benzoylchloride (1.02 ml, 8.89 mmol) was slowly addedat 0° C. under the presence of triethylamine (1.24 ml, 8.89 mmol). Afterstirring reaction solution for 1 hour, the reaction was quenched by 1 Nhydrochloric acid and the reaction solution was extracted withdichloromethane. A pure compound of (E)-1-cyano-benzaldehydeO-benzoyloxime (14) (1.30 g, 81%) was obtained by separating the residuewith silica gel column chromatography.

¹H NMR (300 MHz, CDCl₃) 7.71˜7.50 (m, 6H), 8.04 (m, 2H), 8.25 (m, 2H).

Example 15 Preparation of (E)-1-chloro-benzaldehyde O-benzoyloxime

After dissolving (E)-benzaldehyde oxime (3.0 g, 25 mmol) in CH₂Cl₂ (30ml), N-chlorosuccinimide (3.4 g, 25 mmol) was slowly added at 0° C.After stirring reaction solution for 2 hours, the reaction was completedby adding water (20 ml), and the reaction solution was extracted withdichloromethane. A pure compound of (E)-1-chloro-benzaldehyde oxime(3.30 g, 85%) was obtained by separating the residue with silica gelcolumn chromatography. After dissolving the (E)- and (Z)-oxime mixtures(1.5 g, 9.64 mmol) in CH₂Cl₂ (30 ml), benzoylchloride (1.45 ml, 12.53mmol) was slowly added at 0° C. under the presence of triethylamine(1.75 ml, 12.53 mmol). After stirring reaction solution for 1 hour, thereaction was quenched by 1 N hydrochloric acid, and the reactionsolution was extracted with dichloromethane. A pure compound of(E)-1-chloro-benzaldehyde O-benzoyloxime (15) (1.9 g, 76%) was obtainedby separating the residue with silica gel column chromatography.

¹H NMR (300 MHz, CDCl₃) 7.52 (m, 5H), 7.67 (m, 1H), 8.05 (m, 2H), 8.20(m, 2H).

Example 16 Preparation of (E)-1-benzoic acid-benzaldehyde O-benzoyloxime

After dissolving benzoylchloride (1.0 g, 7.11 mmol) in ethanol (20 ml),hydroxyamine (0.59 g, 8.53 mmol) and triethylamine (1.18 ml, 8.53 mmol)were added. After stirring reaction solution for 30 minutes at roomtemperature, the solvent was removed. After adding water (50 ml), thesolution was extracted with ethylacetate. (E)-1-benzoicacid-benzaldehyde O-benzoyloxime (16) (1.5 g, 61%) was obtained byseparating the residue with column chromatography.

¹H NMR (300 MHz, CDCl₃) 7.29 (m, 2H), 7.43˜7.61 (m, 6H), 7.74 (m, 1H),7.86 (m, 2H), 7.99 (m, 2H), 8.26 (m, 2H).

Example 17 Preparation of (E)-2-furan-2-carboaldehyde O-benzoyloxime

The title compound was prepared in the same method as Example 1 exceptthat 2-furan-2-carbonylaldehyde was used instead of benzaldehyde.

¹H NMR (300 MHz, CDCl₃) 6.55 (dd, J=3.6, 3.6 Hz, 1H), 7.00 (d, J=3.9 Hz,1H), 7.48-7.61 (m, 4H), 8.11 (d, J=2.1, 6.6 Hz, 2H).

Example 18 Preparation of (E)-2-thiophene-2-carboaldehyde O-benzoyloxime

The title compound was prepared in the same method as Example 1 exceptthat 2-thiophene-2-carbonylaldehyde was used instead of benzaldehyde.

¹H NMR (300 MHz, CDCl₃) 7.13 (dd, J=4.8, 4.8 Hz, 1H), 7.46-7.64 (m, 5H),8.10 (dd, J=1.8, 6.6 Hz, 2H), 8.71 (s, 1H).

Example 19 Preparation of (E)-1-phenylethanonaldehyde O-benzoyloxime

The title compound was prepared in the same method as example 1 exceptthat acetophenone was used instead of benzaldehyde.

¹H NMR (300 MHz, CDCl₃) 2.53 (s, 3H), 7.42-7.53 (m, 5H), 7.62 (m, 1H),7.83 (m, 2H), 8.14 (dd, J=1.2, 7.2 Hz, 2H).

Example 20 Preparation of (E)-N-4-fluorobenzylpyrrole-2-carboaldehydeO-benzoyloxime

The title compound was prepared in the same method as example 1 exceptthat N-fluorobenzylpyrrole-2-carbonylaldehyde was used instead ofbenzaldehyde.

¹H NMR (300 MHz, CDCl₃) 5.57 (s, 2H), 6.27 (t, J=3.3 Hz, 1H), 6.63 (dd,J=1.5, 3.9 Hz, 1H), 6.99 (m, 4H), 7.28 (m, 1H), 7.47 (m, 2H), 7.59 (m,1H), 8.07 (m, 2H), 8.37 (s, 1H).

Example 21 Preparation of (E)-3,4-di-fluorobenzaldehyde O-benzoyloxime

The title compound was prepared in the same method as Example 1 exceptthat 3,4-di-fluorobenzaldehyde was used instead of benzaldehyde.

¹H NMR (300 MHz, CDCl₃) 7.25 (m, 1H), 7.50 (m, 3H), 7.63 (m, 1H), 7.73(m, 1H), 8.12 (dd, J=1.8, 9.9 Hz, 2H), 8.50 (s, 1H).

Example 22 Preparation of (E)-3,4-di-fluorobenzaldehydeO-morpholine-4-carbonyloxime

The title compound was prepared in the same method as example 2 exceptthat 3,4-di-fluoroaldehyde was used instead of benzaldehyde.

¹H NMR (300 MHz, CDCl₃) 3.56 (t, J=5.0 Hz, 4H), 3.71 (t, J=4.7 Hz, 4H),7.22 (m, 1H), 7.42 (m, 1H), 7.64 (m, 1H), 8.45 (s, 1H).

Example 23 Preparation of (E)-3,4-di-fluorobenzaldehydeO-4-fluorobenzoyloxime

The title compound was prepared in the same method as Example 1 exceptthat 3,4-di-fluorobenzaldehyde and 4-fluorobenzoylchloride were usedinstead of benzaldehyde and benzoylchloride.

¹H NMR (300 MHz, CDCl₃) 7.17 (m, 2H), 7.26 (m, 1H), 7.52 (m, 1H), 7.72(m, 1H), 8.14 (m, 2H), 8.48 (s, 1H).

Example 24 Preparation of (E)-3,4-di-fluorobenzaldehyde O-oleyloxime

The title compound was prepared in the same method as Example 1 exceptthat 3,4-di-fluorobenzaldehyde and oleoylchloride were used instead ofbenzaldehyde and benzoylchloride.

¹H NMR (300 MHz, CDCl₃) 0.85 (t, J=6.3 Hz, 3H), 1.25 (m, 20H), 1.70 (m,2H), 1.99 (m, 4H), 2.44 (t, J=7.2 Hz, 2H), 5.34 (m, 2H), 7.20 (m, 1H),7.42 (m, 1H), 7.64 (m, 1H), 8.28 (s, 1H).

Example 25 Preparation of (E)-3,4-di-fluorobenzaldehyde O-linoleyloxime

The title compound was prepared in the same method as Example 1 exceptthat 3,4-di-fluorobenzaldehyde and linoleylchloride were used instead ofbenzaldehyde and benzoylchloride.

¹H NMR (300 MHz, CDCl₃) 0.89 (t, J=6.6 Hz, 3H), 1.23-1.41 (m, 14H), 1.73(m, 2H), 2.05 (m, 4H), 2.46 (t, J=7.2 Hz, 2H), 2.77 (t, J=6.3 Hz, 2H),5.35 (m, 4H), 7.22 (m, 1H), 7.44 (m, 1H), 7.65 (m, 1H), 8.29 (s, 1H).

Example 26 Preparation of (E)-3,4-di-fluorobenzaldehyde O-decanoyloxime

The title compound was prepared in the same method as Example 1 exceptthat 3,4-di-fluorobenzaldehyde and decanoylchloride were used instead ofbenzaldehyde and benzoylchloride.

¹H NMR (300 MHz, CDCl₃) 0.88 (t, J=6.6 Hz, 3H), 1.34 (m, 12H), 1.73 (m,2H), 2.47 (t, J=7.2 Hz, 2H), 7.23 (m, 1H), 7.44 (m, 1H), 7.65 (m, 1H),8.29 (s, 1H).

Example 27 Preparation of (E)-3,4-di-fluorobenzaldehydeO-(4-nitro)benzoyloxime

The title compound was prepared in the same method as Example 1 exceptthat 3,4-di-fluorobenzaldehyde and 4-nitrobenzoylchloride were usedinstead of benzaldehyde and benzoylchloride.

¹H NMR (300 MHz, CDCl₃) 7.28 (m, 1H), 7.52 (m, 1H), 7.74 (m, 1H), 8.33(dd, J=9.0, 9.0 Hz, 4H), 8.53 (s, 1H).

Example 28 Preparation of (E)-3,4-di-fluorobenzaldehydeO-3,4-difluorobenzoyloxime

The title compound was prepared in the same method as Example 1 exceptthat 3,4-di-fluorobenzaldehyde and 3,4-difluorobenzoylchloride was usedinstead of benzaldehyde and benzoylchloride.

¹H NMR (300 MHz, CDCl₃) 7.28 (m, 3H), 7.51 (m, 1H), 7.73 (m, 1H), 7.94(m, 2H), 8.49 (s, 1H).

Example 29 Preparation of (E)-2,3-di-fluorobenzaldehyde O-benzoyloxime

The title compound was prepared in the same method as Example 1 exceptthat 2,3-di-fluorobenzaldehyde was used instead of benzaldehyde.

¹H NMR (300 MHz, CDCl₃) 7.16 (m, 1H), 7.29 (m, 1H), 7.50 (t, J=7.7 Hz,2H), 7.63 (m, 1H), 7.88 (m, 1H), 8.13 (m, 2H), 8.82 (s, 1H).

Example 30 Preparation of (E)-2,3-di-fluorobenzaldehydeO-morpholine-4-carbonyloxime

The title compound was prepared in the same method as Example 2 exceptthat 2,3-di-fluorobenzaldehyde was used instead of benzaldehyde.

¹H NMR (300 MHz, CDCl₃) 3.57 (t, J=4.9 Hz, 4H), 3.71 (t, J=4.8 Hz, 4H),7.10 (m, 1H), 7.25 (m, 1H), 7.79 (dd, J=6.2, 8.9 Hz, 1H), 8.57 (s, 1H).

Example 31 Preparation of (E)-2,4-di-fluorobenzaldehyde O-benzoyloxime

The title compound was prepared in the same method as Example 1 exceptthat 2,4-di-fluorobenzaldehyde was used instead of benzaldehyde.

¹H NMR (300 MHz, CDCl₃) 6.88 (m, 1H), 6.97 (m, 1H), 7.50 (m, 2H), 7.61(m, 1H), 8.12 (m, 3H), 8.75 (s, 1H).

Example 32 Preparation of (E)-2,4-di-fluorobenzaldehydeO-morpholine-4-carbonyloxime

The title compound was prepared in the same method as Example 2 exceptthat 2,4-di-fluorobenzaldehyde was used instead of benzoylchloride.

¹H NMR (300 MHz, CDCl₃) 3.55 (t, J=4.8 Hz, 4H), 3.69 (t, J=4.6 Hz, 4H),6.84 (m, 1H), 6.91 (m, 1H), 8.04 (m, 1H), 8.50 (s, 1H).

Example 33 Preparation of (E)-2,4-di-fluorobenzaldehydeO-4-fluorobenzoyloxime

The title compound was prepared in the same method as Example 1 exceptthat 2,4-di-fluorobenzaldehyde and 4-fluorobenzoylchloride were usedinstead of benzaldehyde and benzoylchloride.

¹H NMR (300 MHz, CDCl₃) 6.93 (m, 2H), 7.16 (t, 2H, J=8.6 Hz), 8.15 (m,3H), 8.75 (s, 1H).

Example 34 Preparation of (E)-2,6-di-fluorobenzaldehyde O-benzoyloxime

The title compound was prepared in the same method as Example 1 exceptthat 2,6-di-fluorobenzaldehyde was used instead of benzaldehyde.

¹H NMR (300 MHz, CDCl₃) 7.02 (t, J=8.4 Hz, 2H), 7.47 (m, 3H), 7.62 (t,J=7.2 Hz), 8.12 (m, 1H), 8.76 (s, 1H).

Example 35 Preparation of (E)-2,6-di-fluorobenzaldehydeO-morpholine-4-carbonyloxime

The title compound was prepared in the same method as Example 2 exceptthat 2,6-di-fluorobenzaldehyde was used instead of benzaldehyde.

¹H NMR (300 MHz, CDCl₃) 3.54 (t, J=4.8 Hz, 4H), 3.68 (t, J=4.7 Hz, 4H),7.08 (m, 2H), 7.70 (m, 1H), 8.50 (s, 1H).

Example 36 Preparation of (E)-3,5-di-fluorobenzaldehyde O-benzoyloxime

The title compound was prepared in the same method as example 1 exceptthat 3,5-di-fluorobenzaldehyde was used instead of benzaldehyde.

¹H NMR (300 MHz, CDCl₃) 6.95 (m, 1H), 7.36 (m, 2H), 7.53 (m, 2H), 7.63(m, 1H), 8.12 (m, 2H), 8.50 (s, 1H).

Example 37 Preparation of (E)-3,5-di-fluorobenzaldehydeO-morpholine-4-carbonyloxime

The title compound was prepared in the same method as Example 2 exceptthat 3,5-di-fluorobenzaldehyde was used instead of benzaldehyde.

¹H NMR (300 MHz, CDCl₃) 3.55 (t, J=4.8 Hz, 4H), 3.68 (t, J=4.8 Hz, 4H),6.95 (t, J=8.6 Hz, 2H), 7.38 (m, 1H), 8.48 (s, 1H).

Example 38 Preparation of (E)-2,5-di-fluorobenzaldehydeO-morpholine-4-carbonyloxime

The title compound was prepared in the same method as Example 2 exceptthat 2,5-di-fluorobenzaldehyde was used instead of benzaldehyde.

¹H NMR (300 MHz, CDCl₃) 3.56 (t, J=4.9 Hz, 4H), 3.71 (t, J=4.9 Hz, 4H),6.90 (m, 1H), 7.28 (m, 2H), 8.24 (s, 1H).

Experimental Example 1 The Effect of O-acyloxime Derivatives Accordingto the Present Invention on Lp-PLA₂ Activity

The following tests were carried out to analyze the effect ofO-acyloxime derivatives according to the present invention on Lp-PLA₂activity.

1. Preparation of Enzyme Sources

Blood was collected from normal lipidemic volunteers. EDTA was used asanticoagulant (1.5 mg/ml of blood). After low-speed centrifugation ofthe whole blood to obtain plasma and to prevent lipoproteinmodification, EDTA (0.1%), NaN₃ (0.05%), and PMSF (0.015%) were added.LDL was isolated from the plasma by discontinuous density gradient ultracentrifugation. Briefly, the plasma was centrifuged at 100,000×g at 4°C. for 20 hours. After the top layers containing chylomicron and verylow-density lipoprotein (VLDL) were removed, the density of remainingplasma fractions was increased to 1.064 g/ml with NaBr solution, andthen they were recentrifuged at 100,000×g for an additional 24 hours.The LDL fraction in the top of the tube was collected and dialyzedovernight against three changes of phosphate buffer (pH 7.4), containingNaCl (150 mM), in the dark at 4° C. to remove NaBr and EDTA. The LDL inPBS was stored at 4° C. and used within 4 weeks. The purity of thefraction was confirmed by agarose gel electrophoresis and SDS-PAGE.Concentration of LDL protein was determined using bovine serum albumin(BSA) as a standard.

2. Measurement of Lp-PLA₂ Activity

A method partially modified from the method of Boyd et al. (Bioorg. Med.Chem. Lett., 2000, 10, 2557-2561) was used. Lp-PLA₂, the enzyme is alsoknown as platelet-activating factor acetylhydrolase (PAF-AH), activitywas measured using [³H]PAF as a substrate. Briefly, a micelle substratewas prepared with unlabelled PAF and [³H]PAF (100 μCi/mL, 21.5 Ci/mmole,NET 910) in 10 mM phosphate-buffered saline (PBS), pH 7.4, containing2.7 mM EDTA (PBS-EDTA). The reaction mixture, containing 20 μL ofdiluted human LDL (Lp-PLA₂ source, 4-5 μg protein), 120 μL of PBS-EDTA,and 20 μL of test sample, was preincubated at 37° C. for 15 min. Thereaction was initiated by the addition of 40 μL micelle substrate (0.1μCi, final conc. 80 μM PAF) to measure initial rates of PAF-AH activity.The reaction was stopped by vortexing with 600 μL of CHCl₃/MeOH (2:1)and the CHCl₃ and aqueous layers were separated by centrifugation. Theaqueous layer was removed (250 μL) and vortexed with 250 μL of CHCl₃.The aqueous layer was again removed and the [³H]acetate determined byscintillation counting (1450 Microbeta Trilux, Qallac Oy, Turku,Finland). The raw counts were corrected for background using anonenzyme-containing blank and were expressed as nanomoles of PAFdegraded per hour per milligram of protein.

A portion of the test results is shown in table 2, and figures showaverage values of two test results.

TABLE 2 Sample IC₅₀ (μM)^(a) Example 1

 3.8 Example 7

 4.4 Example 10

11%^(b) Example 11

25.0 Example 16

29%^(b) Example 18

26.0 Example 19

11.2 Example 21

 2.0 Example 24

16%^(b) Example 25

20%^(b) Example 26

12%^(b) Example 27

17%^(b) ^(a)Data indicate average values of two test results.^(b)Inhibition rate at 25 μM

As shown in table 2, O-acyloxime derivatives according to the presentinvention have excellent inhibitory activity of Lp-PLA₂ enzyme.Additionally, another O-acyloxime derivatives according to the presentinvention (not shown in table 2) have showed IC₅₀ value of 0.1 μM˜25 μMand also have showed excellent inhibitory activity.

Accordingly, O-acyloxime derivatives according to the present inventionmay effectively be used for prevention and treatment of cardiovasculardisease caused by Lp-PLA₂, such as hyperlipidemia, coronary arterialheart disease, atherosclerosis, and myocardial infarction.

Experimental Example 2 Acute Toxicity Test of Oral Administration toMouse

The following tests were carried out to observe acute toxicity ofO-acyloxime derivatives according to the present invention.

Specific pathogens free ICR mice of 4 weeks old (12 females and 12males; 3 females and 3 males/dosage group) were raised in an animalchamber controlled with the temperature of 22±3° C., humidity of 55±10%,and illumination of 12L/12D cycle. The mice were acclimated for a weekbefore the test. Feed for a test animal (CJ Corp., for mouse and rat)and water were supplied after sterilization and taken by the micewithout any restriction.

Each 50 mg/ml solution of (E)-3,4-di-fluorobenzaldehyde O-benzoyloximeof Example 21 or (E)-2,3-di-fluorobenzaldehydeO-morpholine-4-carbonyloxime of Example 30 in 0.5% Tween 80 wasprepared. The amount of 0.04 ml (100 mg/kg), 0.2 ml (500 mg/kg), and 0.4ml (1,000 mg/kg) per mouse weight of 20 g was dosed individually to themice by oral administration. The samples were dosed only one time, andadverse effect and mortality were observed during 7 days after thedosing as follows. That is, changes of general symptoms and mortalitywere observed at 1 hour, 4 hours, 8 hours, and 12 hours after theadministration, and one or more times in every morning and afternoonfrom the second day to the seventh day since the dosing day.

Additionally, on seventh day after the administration, the animals weresacrificed and dissected to examine internal organs with naked eye.Weight loss due to (E)-3,4-di-fluorobenzaldehyde O-benzoyloxime or(E)-2,3-di-fluorobenzaldehyde O-morpholine-4-carbonyloxime was observedby measuring weight changes at the intervals of 1 day since the dosage.

According to the test result, no remarkable clinical sympton wasobserved from all the mice dosed with the test material, and no deadmouse was found. Additionally, no toxicity was observed in terms ofweight change, blood test, biochemical test of blood, and necropsyfinding.

Accordingly, (E)-3,4-di-fluorobenzaldehyde O-benzoyloxime and(E)-2,3-di-fluorobenzaldehyde O-morpholine-4-carbonyloxime according tothe present invention did not show any toxicity to all the mice up tothe dosage of 1,000 mg/kg. Therefore, it was identified that thecompounds are safe materials having the lethal dose 50 (LD₅₀) by oraladministration of at least more than 1,000 mg/kg.

Examples of pharmaceutical compositions containing the compoundsaccording to the present invention are described as follows.

Formulation Example 1 Pharmaceutical Formulation

1. Preparation of Powder

O-acyloxime derivative of Formula 1: 2 g Lactose: 1 g

A powder was prepared by mixing the above components and filling theminto an airtight bag.

2. Preparation of Tablet

O-acyloxime derivative of Formula 1: 100 mg Corn starch: 100 mg Lactose:100 mg Magnesium stearate:  2 mg

A tablet was prepared by mixing the above components and tabletting themwith a preparation method of a conventional tablet.

3. Preparation of Capsule

O-acyloxime derivative of Formula 1: 100 mg Corn starch: 100 mg Lactose:100 mg Magnesium stearate  2 mg

A capsule was prepared by mixing the above components and filling theminto a gelatin capsule with a preparation method of a conventionalcapsule.

4. Preparation of Injection Solution

O-acyloxime derivative of Chemical Formula 1: 10 μg/ml, Dilutedhydrochloric acid BP: added until reaching pH 3.5, Sodium chloride BPfor injection: max.1 ml.

After dissolving O-acyloxime derivative of Formula 1 in sodium chlorideBP for injection having a proper volume, pH of the formed solution wascontrolled to pH 3.5 with diluted hydrochloric acid BP. The volume ofthe solution was controlled with sodium chloride BP for injection, andthen sufficiently mixed. After filling the solution into a 5 ml Type Iample made of transparent glass, the ample was sealed by melting theupper empty part of the ample, and sterilized for more than 15 minutesat 120° C. in an autoclave.

O-acyloxime derivatives according to the present invention haveexcellent inhibitory effect on Lp-PLA₂, and may thereby be effectivelyused for prevention and treatment of cardiovascular disease caused byLp-PLA₂ such as hyperlipidemia, coronary arterial heart disease,atherosclerosis, and myocardial infarction.

While this invention has been described in connection with what ispresently considered to be practical example embodiments, it should beunderstood that the invention is not limited to the disclosedembodiments, but, on the contrary, is intended to cover variousmodifications and equivalent arrangements included within the spirit andscope of the appended claims.

1. A compound of O-acyloxime represented by the following Formula 1, or a pharmaceutically acceptable salt thereof:

wherein, R¹ is phenyl, heteroaryl, phenyl or heteroaryl substituted by one or more groups selected from the group consisting of halogen, C₁-C₆ alkyl substituted by one or more halogen, C₆-C₁₀ arylalkyl, C₆-C₁₀ arylalkyl substituted by one or more halogen, and C₆-C₁₀ arylcarbonyl; R² is hydrogen, halogen, cyano, di(C₁-C₃ alkylcarbonyl)amino, or benzoyloxy; and R³ is imidazole, pyrimidine, C₁-C₁₈ alkyl, C₁-C₁₈ alkenyl; cyclohexane; or benzoylamino.
 2. A compound of O-acyloxime represented by the following Formula 1, or a pharmaceutically acceptable salt thereof:

wherein R¹ is phenyl; R² is cyano; and R³ is morpholine.
 3. A compound of O-acyloxime selected from the group consisting of: 5) (E)-benzaldehyde-O-thiophene-2-carbonyloxime, 6) (E)-benzaldehyde-O-furan-2-carbonyloxime, 9) (E)-4-fluorobenzaldehyde O-morpholine-4-carbonyloxime, 22) (E)-3,4-di-fluorobenzaldehyde O-morpholine-4-carbonyloxime, 30) (E)-2,3-di-fluorobenzaldehyde O-morpholine-4-carbonyloxime, 32) (E)-2,4-di-fluorobenzaldehyde O-morpholine-4-carbonyloxime, 35) (E)-2,6-di-fluorobenzaldehyde O-morpholine-4-carbonyloxime, 37) (E)-3,5-di-fluorobenzaldehyde O-morpholine-4-carbonyloxime, and 38) (E)-2,5-di-fluorobenzaldehyde O-morpholine-4-carbonyloxime.
 4. A method for preparing a compound of O-acyloxime of claim 1 comprising: 1) reacting a compound 2 with hydroxyamine in the presence of base to form a compound 3; and 2) reacting the compound 3 prepared in the step 1) with acylchloride to form a compound of O-acyloxime of claim 1, wherein, said compound 2 has a structure of

said compound 3 has a structure of


5. A method for preparing a compound of O-acyloxime of claim 3 comprising: 1) reacting a compound 2 with hydroxyamine in the presence of base to form a compound 3; and 2) reacting the compound 3 prepared in the step 1) with acylchloride to form a compound of O-acyloxime of claim 3, wherein, said compound 2 has a structure of

said compound 3 has a structure of

said compound of O-acyloxime is selected from the group consisting of 5) (E)-benzaldehyde-O-thiophene-2-carbonyloxime, 6) (E)-benzaldehyde-O-furan-2-carbonyloxime, 9) (E)-4-fluorobenzaldehyde O-morpholine-4-carbonyloxime, 22) (E)-3,4-di-fluorobenzaldehyde O-morpholine-4-carbonyloxime, 30) (E)-2,3-di-fluorobenzaldehyde O-morpholine-4-carbonyloxime, 32) (E)-2,4-di-fluorobenzaldehyde O-morpholine-4-carbonyloxime, 35) (E)-2,6-di-fluorobenzaldehyde O-morpholine-4-carbonyloxime, 37) (E)-3,5-di-fluorobenzaldehyde O-morpholine-4-carbonyloxime, and 38) (E)-2,5-di-fluorobenzaldehyde O-morpholine-4-carbonyloxime wherein, R¹ is phenyl, 4-fluorophenyl, 2,3-difluorophenyl, 2,4-difluorophenyl, 2,5-difluorophenyl, 3,4-difluorophenyl, 3,5-difluorophenyl, 2-furan, 2-thiophene, 3-phenylpropenyl, or N-4-fluorobenzylpyrrolyl; and R² is hydrogen amino, benzoyl, chloro, cyano, diacetylamino, or methyl.
 6. A pharmaceutical composition for treatment of cardiovascular disease comprising the compound of claim 1 and pharmaceutically acceptable salts thereof, wherein the cardiovascular disease is selected from the group consisting of hyperlipidemia, coronary arterial heart disease, atherosclerosis, and myocardial infarction.
 7. A Lp-PLA₂ inhibitor comprising the compound of O-acyloxime of claim 1 and pharmaceutically acceptable salts thereof.
 8. A pharmaceutical composition for treatment of cardiovascular disease comprising the compound of claim 2 and pharmaceutically acceptable salts thereof, wherein the cardiovascular disease is selected from the group consisting of hyperlipidemia, coronary arterial heart disease, atherosclerosis, and myocardial infarction.
 9. A pharmaceutical composition for treatment of cardiovascular disease comprising the compound of claim 3 and pharmaceutically acceptable salts thereof, wherein the cardiovascular disease is selected from the group consisting of hyperlipidemia, coronary arterial heart disease, atherosclerosis, and myocardial infarction.
 10. A Lp-PLA₂ inhibitor comprising the compound of O-acyloxime of claim 3 and pharmaceutically acceptable salts thereof.
 11. A compound of O-acyloxime represented by the following Formula 1, or a pharmaceutically acceptable salt thereof:

wherein R¹ is selected from the group consisting of phenyl, heteroaryl, phenyl or heteroaryl substituted by one or more groups selected from the group consisting of halogen, C₁-C₆ alkyl substituted by one or more halogen, C₆-C₁₀ arylalkyl, C₆-C₁₀ arylalkyl substituted by one or more halogen, and C₆-C₁₀ arylcarbonyl; R² is selected from the group consisting of halogen, cyano, di(C₁-C₃ alkylcarbonyl)amino, and benzoyloxy; and R³ is imidazole, pyrimidine, C₂-C₆heterocyclic ring consisting of one or more O or S, C₁-C₁₈alkyl, C₁-C₁₈ alkenyl; cyclohexane; or benzoylamino.
 12. A Lp-PLA₂ inhibitor comprising the compound of O-acyloxime of claim 11 and pharmaceutically acceptable salts thereof. 